The emergence of metal oxide nanoparticles (NPs) as a phytomedicine: A two-facet role in plant growth, nano-toxicity and anti-phyto-microbial activity.

Anti-microbial resistance (AMR) has recently emerged as an area of high interest owing to the rapid surge of AMR phenotypes. Metal oxide NPs (MeONPs) have been identified as novel phytomedicine and have recently peaked a lot of interest due to their potential applications in combating phytopathogens, besides enhancing plant growth and yields. Numerous MeONPs (Ti2O, MgO, CuO, Ag2O, SiO2, ZnO, and CaO) have been synthesized and tested to validate their antimicrobial roles without causing toxicity to the cells. This review discusses the application of the MeONPs with special emphasis on anti-microbial activities in agriculture and enlists how cellular toxicity caused through reactive oxygen species (ROS) production affects plant growth, morphology, and viability. This review further highlights the two-facet role of silver and copper oxide NPs including their anti-microbial applications and toxicities. Furthermore, the factor modulating nanotoxicity and immunomodulation for cytokine production has also been discussed. Thus, this article will not only provide the researchers with the potential bottlenecks but also emphasizes a comprehensive outline of breakthroughs in the applicability of MeONPs in agriculture.

Contracaecum nematode parasites in hillstream loaches of the Western Ghats, India.

Nematode parasites of the family Anisakidae infect definitive hosts, such as fish-eating birds and mammals, through primary intermediate hosts like copepods and secondary intermediate hosts like fishes. However, consumption of raw or undercooked fish can lead to nematode infection called anisakidosis in humans. We observed the presence of nematode infection in hillstream loaches of families Cobitidae and Nemacheilidae available for human consumption in the local markets in the northern parts of Western Ghats, India. Scanning electron micrograph and genetic identification employing mitochondrial cytochrome oxidase subunit II, identified the nematode to the genus Contracaecum. Histology of infected host revealed the presence of the parasite in muscles. Antioxidant enzyme analysis of host liver suggested that infection leads to oxidative stress in the fish. We suspect that a gradual increase in parasite infection of the loaches in the last decade could be attributed to various anthropogenic stressors that are altering riverine habitats. Since loaches are consumed by tribal people who often prepare the fish without degutting and possibly undercooked, there is a potential threat of human infection.

Micro-contaminant, but immense impact: Source and influence of diethyl phthalate plasticizer on bottom-dwelling fishes.

Sustainable plastic-waste management is becoming increasingly challenging as enormous loads of plastic debris regularly accumulate in susceptible ecosystems. The microplastic (MP) particles generated from these plastic wastes are imposing additional threats to these ecosystems due to their small size as well as their ability to adsorb and carry toxic chemicals. The current investigation deals with one such MP-originated toxicant, diethyl phthalate (DEP), and its impact on two species of freshwater loaches from the Western Ghats of India, Lepidocephalichthys thermalis and Indoreonectes evezardi. The MP samples were collected from the sediments of the Mula River and characterized using spectroscopic methods and scanning electron microscopy. Polymers, such as polyvinyl chloride and polypropylene, were identified in the collected MPs. GC-MS analysis of the MP extracts revealed the presence of DEP, confirming the MP waste as a potential source of DEP pollution. Further, to evaluate the effect of DEP on survival of selected loaches, L. thermalis and I. evezardi were exposed to DEP concentrations (18.75-300 mg L-1) and the lethal DEP dose (LC50) was estimated to be 44.53 mg L-1 for L. thermalis and 34.64 mg L-1 for I. evezardi. Fishes were further exposed to sub-lethal DEP concentration for one day (Short term exposure: STE) or eight days (Long term exposure: LTE) to analyze the histological condition and oxidative status of the liver in response to DEP treatment. Histology revealed congestion of sinusoids and vacuolization after the LTE. Higher lipid peroxidation levels were also measured in the livers of both species treated with DEP, which indicated DEP-mediated oxidative damage. The antioxidant enzymes including superoxide dismutase, catalase, peroxidase and glutathione-S-transferase displayed elevated activities in response to STE and LTE of DEP. Collectively, the results demonstrate that MPs in the Mula River are a potential source of DEP. The findings also show that DEP exposure can be fatal to freshwater fishes such as loaches, possibly by causing increased oxidative damage to the hepatic system.

Exploring Phytochemicals for Combating Antibiotic Resistance in Microbial Pathogens.

Antibiotic resistance or microbial drug resistance is emerging as a serious threat to human healthcare globally, and the multidrug-resistant (MDR) strains are imposing major hurdles to the progression of drug discovery programs. Newer antibiotic-resistance mechanisms in microbes contribute to the inefficacy of the existing drugs along with the prolonged illness and escalating expenditures. The injudicious usage of the conventional and commonly available antibiotics in human health, hygiene, veterinary and agricultural practices is proving to be a major driver for evolution, persistence and spread of antibiotic-resistance at a frightening rate. The drying pipeline of new and potent antibiotics is adding to the severity. Therefore, novel and effective new drugs and innovative therapies to treat MDR infections are urgently needed. Apart from the different natural and synthetic drugs being tested, plant secondary metabolites or phytochemicals are proving efficient in combating the drug-resistant strains. Various phytochemicals from classes including alkaloids, phenols, coumarins, terpenes have been successfully demonstrated their inhibitory potential against the drug-resistant pathogens. Several phytochemicals have proved effective against the molecular determinants responsible for attaining the drug resistance in pathogens like membrane proteins, biofilms, efflux pumps and bacterial cell communications. However, translational success rate needs to be improved, but the trends are encouraging. This review highlights current knowledge and developments associated challenges and future prospects for the successful application of phytochemicals in combating antibiotic resistance and the resistant microbial pathogens.

Nitric oxide, crosstalk with stress regulators and plant abiotic stress tolerance.

KEY MESSAGE: Nitric oxide is a dynamic gaseous molecule involved in signalling, crosstalk with stress regulators, and plant abiotic-stress responses. It has great exploratory potentials for engineering abiotic stress tolerance in crops. Nitric oxide (NO), a redox-active gaseous signalling molecule, though present uniformly through the eukaryotes, maintain its specificity in plants with respect to its formation, signalling, and functions. Its cellular concentrations are decisive for its function, as a signalling molecule at lower concentrations, but triggers nitro-oxidative stress and cellular damage when produced at higher concentrations. Besides, it also acts as a potent stress alleviator. Discovered in animals as neurotransmitter, NO has come a long way to being a stress radical and growth regulator in plants. As a key redox molecule, it exhibits several key cellular and molecular interactions including with reactive chemical species, hydrogen sulphide, and calcium. Apart from being a signalling molecule, it is emerging as a key player involved in regulations of plant growth, development and plant-environment interactions. It is involved in crosstalk with stress regulators and is thus pivotal in these stress regulatory mechanisms. NO is getting an unprecedented attention from research community, being investigated and explored for its multifaceted roles in plant abiotic stress tolerance. Through this review, we intend to present the current knowledge and updates on NO biosynthesis and signalling, crosstalk with stress regulators, and how biotechnological manipulations of NO pathway are leading towards developing transgenic crop plants that can withstand environmental stresses and climate change. The targets of various stress responsive miRNA signalling have also been discussed besides giving an account of current approaches used to characterise and detect the NO.

Embelin-loaded chitosan gold nanoparticles interact synergistically with ciprofloxacin by inhibiting efflux pumps in multidrug-resistant Pseudomonas aeruginosa and Escherichia coli.

A global upsurge in emergence and spread of antibiotic resistance (ABR) in bacterial populations is a serious threat for human health. Unfortunately, ABR is no longer confined to nosocomial environments and is frequently reported from community microbes as well. The ABR is resulting in shrinking potent antibiotics pool and thus necessitating novel and alternative therapies and therapeutics. Current investigation was aimed to assess the synergistic potential of a synthesized, phytomolecule-loaded, polysaccharide-stabilized metallic nanoparticles (NPs) against Pseudomonas aeruginosa (PA) and Escherichia coli (EC) isolated from river waters. ABR profiling of these strains characterized them as multidrug resistant (MDR). Synthesized embelin (Emb, isolated from Embelia tsjeriam-cottam)-loaded, chitosan-gold (Emb-Chi-Au) NPs were assessed for their potential synergistic activity with ciprofloxacin (CIP) via checker-board assay and time-kill curve analysis. The NPs reduced the minimal inhibitory concentration (MIC) of CIP by 16- and 4-fold against MDR PA (PA-r) and EC (EC-r) strains, respectively. Fractional inhibitory concentration (FIC) indices with ≤0.5 values confirmed the synergy between the Emb-Chi-Au NPs and CIP, which was further confirmed at ½ MICs in both PA-r and EC-r via time-kill curve analysis. In order to decipher the mode of action, efflux pump inhibitory effects of Emb-Chi-Au NPs were evaluated in terms of the increase in the EtBr mediated fluorescence in control versus NP-treated MDR strains. Molecular docking based in silico simulations were used to predict the interactions between Emb and the active sites of the efflux pump related proteins in PA-r (MexA, MexB and OprM) and EC-r (AcrA, AcrB and TolC), which revealed the probable bond formation between Emb and respective amino acid residues.

Plant synthetic biology for producing potent phyto-antimicrobials to combat antimicrobial resistance.

Inappropriate and injudicious use of antimicrobial drugs in human health, hygiene, agriculture, animal husbandry and food industries has contributed significantly to rapid emergence and persistence of antimicrobial resistance (AMR), one of the serious global public health threats. The crisis of AMR versus slower discovery of newer antibiotics put forth a daunting task to control these drug-resistant superbugs. Several phyto-antimicrobials have been identified in recent years with direct-killing (bactericidal) and/or drug-resistance reversal (re-sensitization of AMR phenotypes) potencies. Phyto-antimicrobials may hold the key in combating AMR owing to their abilities to target major microbial drug-resistance determinants including cell membrane, drug-efflux pumps, cell communication and biofilms. However, limited distribution, low intracellular concentrations, eco-geographical variations, beside other considerations like dynamic environments, climate change and over-exploitation of plant-resources are major blockades in full potential exploration phyto-antimicrobials. Synthetic biology (SynBio) strategies integrating metabolic engineering, RNA-interference, genome editing/engineering and/or systems biology approaches using plant chassis (as engineerable platforms) offer prospective tools for production of phyto-antimicrobials. With expanding SynBio toolkit, successful attempts towards introduction of entire gene cluster, reconstituting the metabolic pathway or transferring an entire metabolic (or synthetic) pathway into heterologous plant systems highlight the potential of this field. Through this perspective review, we are presenting herein the current situation and options for addressing AMR, emphasizing on the significance of phyto-antimicrobials in this apparently post-antibiotic era, and effective use of plant chassis for phyto-antimicrobial production at industrial scales along with major SynBio tools and useful databases. Current knowledge, recent success stories, associated challenges and prospects of translational success are also discussed.

Genome-wide identification, characterization and transcriptional profiling of NHX-type (Na+/H+) antiporters under salinity stress in soybean.

This study was aimed at the genome-wide identification, a comprehensive in silico characterization of NHX genes from soybean (Glycine max L.) and their tissue-specific expression under varied levels (0-200 mM NaCl) of salinity stress. A total of nine putative NHX genes were identified from soybean. The phylogenetic analysis confirmed a total of five sub-groups and GmNHXs were distributed in three of them. Bioinformatics analyses confirmed all GmNHXs as ion transporters in nature, and all were localized on the vacuolar membrane. Several cis-acting regulatory elements involved in hormonal signal-responsiveness and abiotic stress including salinity responses were identified in the promoter regions of GmNHXs. Amiloride, which is a known Na+/H+ exchanger activity inhibitor, binding motifs were observed in all the GmNHXs. Furthermore, the identified GmNHXs were predicted-targets of 75 different miRNA candidates. To gain an insight into the functional divergence of GmNHX transporters, qRT-PCR based gene expression analysis was done in control and salt-treated root, stem and leaf tissues of two contrasting Indian soybean varieties MAUS-47 (tolerant) and Gujosoya-2 (sensitive). The gene up-regulation was tissue-specific and varied amongst the soybean varieties, with higher induction in tolerant variety. Maximum induction was observed in GmNHX2 in root tissues of MAUS-47 at 200 mM NaCl stress. Overall, identified GmNHXs may be explored further as potential gene candidates for soybean improvement.

Prospects of Exploring the Metal-Organic Framework for Combating Antimicrobial Resistance.

Infectious diseases are a major public health concern globally. Infections caused by pathogens with resistance against commonly used antimicrobial drugs or antibiotics (known as antimicrobial resistance, AMR) are becoming extremely difficult to control. AMR has thus been declared as one of the top 10 global public health threats, as it has very limited solutions. The drying pipeline of effective antibiotics has further worsened the situation. There is no absolute treatment, and the limitations of existing methods warrant further development in antimicrobials. Recent developments in the nanomaterial field present them as promising therapeutics and effective alternative to conventional antibiotics and synthetic drugs. The metal-organic framework (MOF) is a recent addition to the antimicrobial category with superior properties. The MOF exerts antimicrobial action on a wide range of species and is highly biocompatible. Additionally, their porous structures allow the incorporation of biomolecules and drugs for synergistic antimicrobial action. This review provides an inclusive summary of the molecular events responsible for resistance development and current trends in antimicrobials to combat antibiotic resistance and explores the potential role of the MOF in tackling the drug-resistant microbial species.

Individual and additive stress impacts of Na+ and Cl‾ on proline metabolism and nitrosative responses in rice.

It is well-established that plants accumulate high concentrations of sodium (Na+) and chloride (Cl‾) ions when subjected to salinity stress. However, little is known about individual or relative toxic impacts of these ions and whether they exert additive impacts under NaCl. Most of the investigations have historically been directed to decode Na+-toxicity, and as a result deeper understandings about Cl‾-toxicity are lacking. In this study, the extent to which sodium and chloride ions contribute in inducing nitrosative responses and proline metabolism is shown in two rice cultivars, one tolerant (Panvel-3) and one sensitive (Sahyadri-3). Equimolar (100 mM) concentrations of Na+, Cl‾ and NaCl (EC ≈ 10 dSm-1) reduced biomass production in both the cultivars in following manner NaCl > Na+>Cl‾. Na+ and NaCl treatments displaced K+, however, the tolerant cultivar maintained low Na+/K+ levels. Hyper-accumulation of Na+ may apparently be attributed for the reduced plant growth and biomass accumulation, and higher lipid-peroxidation. Nitric oxide and nitrate reductase were more responsive to NaCl followed by Na+ and Cl‾, respectively. The expression patterns of key-genes involved in proline biosynthesis and degradation confirmed the involvement of proline in better performance of salt tolerant cultivar under stresses, with higher responsiveness to NaCl and then Na+ and Cl‾ treatments. Principal component analysis revealed correlations in proline metabolism and nitrosative responses under ionic stresses and confirmed the closeness of NaCl and Na+ stresses. Overall, amongst the individual ions, Na+ induced higher toxicity than Cl‾ and both these ions exerted additive stress impacts under NaCl treatment.

Biotic elicitors enhance diosgenin production in Helicteres isora L. suspension cultures via up-regulation of CAS and HMGR genes.

In an attempt to find an alternative and potent source of diosgenin, a steroidal saponin in great demand for its pharmaceutical importance, Helicteres isora suspension cultures were explored for diosgenin extraction. The effect of biotic elicitors on the biosynthesis of diosgenin, in suspension cultures of H. isora was studied. Bacterial as well as fungal elicitors such as Escherichia coli, Bacillus subtilis, Saccharomyces cerevisiae and Aspergillus niger were applied at varying concentrations to investigate their effects on diosgenin content. The HPLC based quantification of the treated samples proved that amongst the biotic elicitors, E. coli (1.5%) proved best with a 9.1-fold increase in diosgenin content over respective control cultures. Further, the scaling-up of the suspension culture to shake-flask and ultimately to bioreactor level were carried out for production of diosgenin. During all the scaling-up stages, diosgenin yield obtained was in the range between 7.91 and 8.64 mg l-1, where diosgenin content was increased with volume of the medium. The quantitative real-time PCR (qRT-PCR) analysis showed biotic elicitors induced the expression levels of regulatory genes in diosgenin biosynthetic pathway, the 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) and cycloartenol synthase (CAS), which can be positively correlated with elicited diosgenin contents in those cultures. The study holds significance as H. isora represents a cleaner and easy source of diosgenin where unlike other traditional sources, it is not admixed with other steroidal saponins, and the scaled-up levels of diosgenin achieved herein have the potential to be explored commercially.

Engineering salinity tolerance in plants: progress and prospects.

MAIN CONCLUSION: There is a need to integrate conceptual framework based on the current understanding of salt stress responses with different approaches for manipulating and improving salt tolerance in crop plants. Soil salinity exerts significant constraints on global crop production, posing a serious challenge for plant breeders and biotechnologists. The classical transgenic approach for enhancing salinity tolerance in plants revolves by boosting endogenous defence mechanisms, often via a single-gene approach, and usually involves the enhanced synthesis of compatible osmolytes, antioxidants, polyamines, maintenance of hormone homeostasis, modification of transporters and/or regulatory proteins, including transcription factors and alternative splicing events. Occasionally, genetic manipulation of regulatory proteins or phytohormone levels confers salinity tolerance, but all these may cause undesired reduction in plant growth and/or yields. In this review, we present and evaluate novel and cutting-edge approaches for engineering salt tolerance in crop plants. First, we cover recent findings regarding the importance of regulatory proteins and transporters, and how they can be used to enhance salt tolerance in crop plants. We also evaluate the importance of halobiomes as a reservoir of genes that can be used for engineering salt tolerance in glycophytic crops. Additionally, the role of microRNAs as critical post-transcriptional regulators in plant adaptive responses to salt stress is reviewed and their use for engineering salt-tolerant crop plants is critically assessed. The potentials of alternative splicing mechanisms and targeted gene-editing technologies in understanding plant salt stress responses and developing salt-tolerant crop plants are also discussed.

The alarming antimicrobial resistance in ESKAPEE pathogens: Can essential oils come to the rescue?

Antibiotics, considered as a backbone of modern clinical-medicines, are facing serious threats from emerging antimicrobial-resistance (AMR) in several bacteria from nosocomial and community origins and is posing a serious human-health concern. Recent commitment by the Heads of States at the United Nations General Assembly (UNGA, 2016) for coordinated efforts to curb such infections illustrates the scale of this problem. Amongst the drug-resistant microbes, major threat is posed by the group named as ESKAPEE, an acronym for Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp., and Escherichia coli, comprising high to critical drug-resistant, World Health Organization Critical Priority I and II pathogens. The drying pipeline of effective and new antibiotics has worsened the situation with looming threat of heading to a 'post-antibiotic era'. This necessitates novel and effective approaches to combat this life-threatening issue. Medicinal and aromatic plants are hailed as the reservoir of bioactive compounds and can serve as a source of antimicrobial compounds, and some recent leads show that essential oils (EOs) may provide an effective solution for tackling AMR. EOs have shown wide-spectrum antimicrobial potentials via targeting the major determinants of pathogenicity, drug-resistance and its spread including cell membrane, drug efflux pumps, quorum sensing, biofilms and R-plasmids. Latest reports confirm the EOs having strong direct-killing or re-sensitizing potentials to replace or rejuvenate otherwise fading antibiotics arsenal. We discuss herein possibilities of using EOs directly for antimicrobial potentials or in combination with antibiotics to potentiate the later for combating AMR in ESKAPEE pathogens. The current understandings, success stories and challenges for translational success have also been discussed.

Phytochemical profile, anti-oxidant, anti-inflammatory, and anti-proliferative activities of Pogostemon deccanensis essential oils.

Essential oils (EOs) obtained from aerial parts of Pogostemon deccanensis were analyzed for GC-MS profiling, and evaluated for antioxidant, anti-inflammatory, and anti-proliferative activities. GC-MS analysis revealed a total of 47 constituents, establishing the EOs rich in sesquiterpene with > 20 sesquiterpenes constituting around 77% of the total EO yield. Major constituents included Curzerene (Benzofuran, 6-ethenyl-4,5,6,7-tetrahydro-3,6-dimethyl-5-isopropenyl-, trans-) (26.39%) and epi-Cadinol (22.68%), Ethanone, 1-(2,4,6-trihydroxyphenyl) (6.83%, Acetophenones), and Boldenone (3.47%, anabolic steroid). EOs found to be rich in phytochemicals attributed for antioxidant potentials of aromatic/medicinal plants, viz., flavonoids (2.71 µg quercetin equivalents g-1 EO), total phenols (3.94 µg gallic acid equivalents (GAE) g-1 EO), carotenoids (14.3 µg ß-carotene equivalents g-1 EO), and ascorbic acid (2.21 µg ascorbic acid equivalents g-1 EO). P. deccanensis EOs exhibited striking antioxidant activities assessed by wide range of assays including ferric reducing antioxidant potential (FRAP, 255.3 GAE at 2 µg mL-1 EO), total antioxidant activity (TAA, 264.3 GAE at 2 µg ml-1) of EO, DPPH (65% inhibition at 2 µg mL-1), and OH (58% inhibition at 2 µg mL-1) scavenging. Interestingly, EOs showed considerably higher anti-lipid peroxidation activity than the standard antioxidant molecule ascorbic acid, with 50% protection by 1.29 µg mL-1 EO against 20.0 µg mL-1 standard. EOs showed strong anti-inflammatory activity with 50% inhibition at 1.95 µg mL-1 EO. The anti-proliferative activity of EOs was tested against mouse cancer cell line and the EOs proved a potent anti-proliferative agent with only 2.1% cell survival at 2 µg mL-1 EO, whereas the EOs were largely non-toxic-to-normal (non-cancerous) cells with approximately 80% cell survival at the 2 µg mL-1 EOs. This being the first attempt of phytochemical profiling and wide array of biological activities of P. deccanensis EOs holds significance as the striking activities were observed at very low concentrations, in some cases at lower than the commercial standards, and has, therefore, great potential for pharmaceutical or commercial exploration.

Exploring miRNAs for developing climate-resilient crops: A perspective review.

Climate changes and environmental stresses have significant implications on global crop production and necessitate developing crops that can withstand an array of climate changes and environmental perturbations such as irregular water-supplies leading to drought or water-logging, hyper soil-salinity, extreme and variable temperatures, ultraviolet radiations and metal stress. Plants have intricate molecular mechanisms to cope with these dynamic environmental changes, one of the most common and effective being the reprogramming of expression of stress-responsive genes. Plant microRNAs (miRNAs) have emerged as key post-transcriptional and translational regulators of gene-expression for modulation of stress implications. Recent reports are establishing their key roles in epigenetic regulations of stress/adaptive responses as well as in providing plants genome-stability. Several stress responsive miRNAs are being identified from different crop plants and miRNA-driven RNA-interference (RNAi) is turning into a technology of choice for improving crop traits and providing phenotypic plasticity in challenging environments. Here we presents a perspective review on exploration of miRNAs as potent targets for engineering crops that can withstand multi-stress environments via loss-/gain-of-function approaches. This review also shed a light on potential roles plant miRNAs play in genome-stability and their emergence as potent target for genome-editing. Current knowledge on plant miRNAs, their biogenesis, function, their targets, and latest developments in bioinformatics approaches for plant miRNAs are discussed. Though there are recent reviews discussing primarily the individual miRNAs responsive to single stress factors, however, considering practical limitation of this approach, special emphasis is given in this review on miRNAs involved in responses and adaptation of plants to multi-stress environments including at epigenetic and/or epigenomic levels.

Plant small RNAs: the essential epigenetic regulators of gene expression for salt-stress responses and tolerance.

Saline environment cues distort the plant growth, development and crop yield. Epigenetics has emerged as one of the prime themes in plant functional genomics for molecular-stress-physiology research, as copious studies have provided new visions into the epigenetic control of stress adaptations. The epigenetic control is associated with the regulation of the expression of stress-related genes which also comprises many steady alterations inherited in next cellular generation as stress memory. These epigenetic amendments also implicate induction of small RNA (sRNA)-mediated fine-tuning of transcriptional and post-transcriptional regulations of gene expression. These tiny (19-24 nt) RNA species, particularly microRNAs (miRNAs) besides endogenous small interfering RNA (siRNA) have emerged as important responsive entities for epigenetic modulation of salt-stress effects on plants. There is a recent upsurge in development of tools and databases useful for prediction, identification and validation of small RNAs (sRNAs) and their target messenger RNAs (mRNAs). Therefore, these small but key regulatory molecules have received a wide attention in post-genomic era as potential targets for engineering stress tolerance in major glycophytic crops, though it is yet to be explored optimally. This review aims to provide critical updates on plant sRNAs as key epigenetic regulators of plant salt-stress responses, their target prediction and validation, computational tools and databases available for plant small RNAs, besides discussing their roles in salt-stress regulatory networks and adaptive mechanisms in plants, with special emphasis on their exploration for engineering salinity tolerance in plants.

Engineering Crops for the Future: A Phosphoproteomics Approach.

Abiotic stresses like salinity, drought, heat, metal ions, radiation and oxidative stress, and especially their combinations, are major limiting factors for growth and productivity of the crops. Various molecular and biochemical processes governing the plant responses to abiotic stresses have often been investigated and hold the key for producing high-yielding and abiotic stress-tolerant crops. Plant responses to abiotic stresses are dynamic and intricate, and vary with type, level, and duration of the stress involved, as well as on the type of tissue under stress. However, one biochemical indicator common to all stresses is definite and controlled protein phosphorylation which is generally transmitted by highly complex protein kinase cascades. In recent years, using different biochemical as well as computational tools, many of such phosphoproteins are identified and characterized with respect to abiotic stresses. Subsequently, an upsurge has been witnessed in recent times for phosphoproteomics repositories or databases. The use of this crucial knowledge about such proteins and their phosphorylation sites is one of the promising ways for crop engineering against abiotic stress. Several reports have described abiotic stress-induced transcriptome, proteome and phosphoproteome changes in plants subjected to these stress factors. However, the investigations to assess precise phosphoproteomics deviations in response to environmental stresses and their implementation for crop improvement are limited. The present review summarizes and discusses the recent developments in deciphering abiotic stress induced changes in plant phosphoproteome besides development of phosphoproteomics tools and their repositories. A critical assessment of targeting phosphoproteins for crop improvement and phosphoproteomics mediated enhanced abiotic stress tolerance in transgenic plants has been presented.

Inhibiting Bacterial Drug Efflux Pumps via Phyto-Therapeutics to Combat Threatening Antimicrobial Resistance.

Antibiotics, once considered the lifeline for treating bacterial infections, are under threat due to the emergence of threatening antimicrobial resistance (AMR). These drug-resistant microbes (or superbugs) are non-responsive to most of the commonly used antibiotics leaving us with few treatment options and escalating mortality-rates and treatment costs. The problem is further aggravated by the drying-pipeline of new and potent antibiotics effective particularly against the drug-resistant strains. Multidrug efflux pumps (EPs) are established as principal determinants of AMR, extruding multiple antibiotics out of the cell, mostly in non-specific manner and have therefore emerged as potent drug-targets for combating AMR. Plants being the reservoir of bioactive compounds can serve as a source of potent EP inhibitors (EPIs). The phyto-therapeutics with noteworthy drug-resistance-reversal or re-sensitizing activities may prove significant for reviving the otherwise fading antibiotics arsenal and making this combination-therapy effective. Contemporary attempts to potentiate the antibiotics with plant extracts and pure phytomolecules have gained momentum though with relatively less success against Gram-negative bacteria. Plant-based EPIs hold promise as potent drug-leads to combat the EPI-mediated AMR. This review presents an account of major bacterial multidrug EPs, their roles in imparting AMR, effective strategies for inhibiting drug EPs with phytomolecules, and current account of research on developing novel and potent plant-based EPIs for reversing their AMR characteristics. Recent developments including emergence of in silico tools, major success stories, challenges and future prospects are also discussed.

MicroRNAs As Potential Targets for Abiotic Stress Tolerance in Plants.

The microRNAs (miRNAs) are small (20-24 nt) sized, non-coding, single stranded riboregulator RNAs abundant in higher organisms. Recent findings have established that plants assign miRNAs as critical post-transcriptional regulators of gene expression in sequence-specific manner to respond to numerous abiotic stresses they face during their growth cycle. These small RNAs regulate gene expression via translational inhibition. Usually, stress induced miRNAs downregulate their target mRNAs, whereas, their downregulation leads to accumulation and function of positive regulators. In the past decade, investigations were mainly aimed to identify plant miRNAs, responsive to individual or multiple environmental factors, profiling their expression patterns and recognizing their roles in stress responses and tolerance. Altered expressions of miRNAs implicated in plant growth and development have been reported in several plant species subjected to abiotic stress conditions such as drought, salinity, extreme temperatures, nutrient deprivation, and heavy metals. These findings indicate that miRNAs may hold the key as potential targets for genetic manipulations to engineer abiotic stress tolerance in crop plants. This review is aimed to provide recent updates on plant miRNAs, their biogenesis and functions, target prediction and identification, computational tools and databases available for plant miRNAs, and their roles in abiotic stress-responses and adaptive mechanisms in major crop plants. Besides, the recent case studies for overexpressing the selected miRNAs for miRNA-mediated enhanced abiotic stress tolerance of transgenic plants have been discussed.

Na+ and Cl(-) ions show additive effects under NaCl stress on induction of oxidative stress and the responsive antioxidative defense in rice.

Despite the fact that when subjected to salinity stress most plants accumulate high concentrations of sodium (Na(+)) and chloride (Cl(-)) ions in their tissues, major research has however been focused on the toxic effects of Na(+). Consequently, Cl(-) toxicity mechanisms in annual plants, particularly in inducing oxidative stress, are poorly understood. Here, the extent to which Na(+) and/or Cl(-) ions contribute in inducing oxidative stress and regulating the adaptive antioxidant defense is shown in two Indica rice genotypes differing in their salt tolerance. Equimolar (100 mM) concentrations of Na(+), Cl(-), and NaCl (EC ≈ 10 dS m(-1)) generated free-radical (O2 (•-), (•)OH) and non-radical (H2O2) forms of reactive oxygen species (ROS) and triggered cell death in leaves of 21-day-old hydroponically grown rice seedlings as evident by spectrophotometric quantifications and histochemical visualizations. The magnitude of ROS-mediated oxidative damage was higher in sensitive cultivar, whereas NaCl proved to be most toxic among the treatments. Salt treatments significantly increased activities of antioxidant enzymes and their isozymes including superoxide dismutase, catalase, peroxidase, ascorbate peroxidase, and glutathione reductase. Na(+) and Cl(-) ions showed additive effects under NaCl in activating the antioxidant enzyme machinery, and responses were more pronounced in tolerant cultivar. The expression levels of SodCc2, CatA, and OsPRX1 genes were largely consistent with the activities of their corresponding enzymes. Salt treatments caused an imbalance in non-enzymatic antioxidants ascorbic acid, α-tocopherol, and polyphenols, with greater impacts under NaCl than Na(+) and Cl(-) separately. Results revealed that though Cl(-) was relatively less toxic than its counter-cation, its effects cannot be totally ignored. Both the cultivars responded in the same manner, but the tolerant cultivar maintained lower Na(+)/K(+) and ROS levels coupled with better antioxidant defense under all three salt treatments.